Localization of Intracellular Antigens in thin Sections with Ferritin Labeled Antibody

1970 ◽  
Vol 28 ◽  
pp. 174-175
Author(s):  
M.S. Shahrabadi ◽  
T. Yamamoto
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Antigenic Determinants ◽  
Conjugate Prior ◽  
Thin Sections ◽  
Specific Attachment ◽  
Intracellular Antigen ◽  
Antigen Antibody ◽  
Ideal Method ◽  
The Ideal

The technique of labeling of macromolecules with ferritin conjugated antibody has been successfully used for extracellular antigen by means of staining the specimen with conjugate prior to fixation and embedding. However, the ideal method to determine the location of intracellular antigen would be to do the antigen-antibody reaction in thin sections. This technique contains inherent problems such as the destruction of antigenic determinants during fixation or embedding and the non-specific attachment of conjugate to the embedding media. Certain embedding media such as polyampholytes (2) or cross-linked bovine serum albumin (3) have been introduced to overcome some of these problems.

scholarly journals THE NATURE OF ANTIGEN-ANTIBODY COMPLEXES FORMED IN RABBITS DURING AN IMMUNE RESPONSE TO BOVINE SERUM ALBUMIN

1958 ◽  
Vol 107 (5) ◽  
pp. 653-663 ◽  
Author(s):  
William O. Weigle
Keyword(s):  
Immune Response ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Ammonium Sulfate ◽  
Bovine Serum ◽  
Ammonium Sulfate Precipitation ◽  
Antibody Synthesis ◽  
Circulating Antigen ◽  
Antigen Antibody

The immune elimination of soluble BSA, following an intravenous injection, is accompanied by the appearance of circulating antigen-antibody complexes. The pattern of the appearance of circulating antigen-antibody complexes and the immune elimination of antigen probably depends on the amount of antigen injected, the rate of antibody synthesis, and perhaps, the quality of antibody produced. There is no relationship between the I* antigen-antibody complexes detected during the immune response in rabbits by ammonium sulfate precipitation and the material precipitated from immune sera as a result of treatment with alkali. Alkali-precipitable material present in the serum of rabbits at a time when I* antigen is also present contain at most only traces of the antigen.

ANTI-BOVINE SERUM ALBUMIN AND ANTI-ALPHA LACTALBUMIN IN THE SERUM OF CHILDREN AND ADULTS

PEDIATRICS ◽  
1965 ◽  
Vol 35 (4) ◽  
pp. 571-588
Author(s):  
Richard M. Rothberg ◽  
Richard S. Farr
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Gamma Globulin ◽  
Age Distribution ◽  
Skin Tests ◽  
Positive Skin ◽  
Antigen Antibody ◽  
Alpha Lactalbumin

1. Because precipitin, hemagglutination, and complement-fixation tests measure secondary manifestations of antigen-antibody interactions and are sometimes negative even after a primary antigen-antibody reaction has occurred in vitro, the incidence and amount of anti-bovine serum albumin (BSA) was evaluated in the sera from 900 children and adults by means of precipitating I131-labeled BSA-antibody complexes with 50% saturated ammonium sulfate. A similar study using I131-labeled alpha lactalbumin (ALA) was performed on 718 of this same group of sera. 2. Antibody to BSA was detected more frequently among children (75%) than among young adults 16 to 40 years of age (25%), or among older age groups (8%). 3. The incidence of detectable antibody to ALA had the same age distribution, but only half the frequency as anti-BSA. 4. In contrast to the near absence of antibody in the cord serum as measured by hemagglutination titers using red cells coated with milk proteins, most of the antibody detected in maternal sera in the present study was able to cross the placental barrier and was present in the cord sera. 5. The incidence of both anti-BSA or anti-ALA was the same in males and females. 6. If a given sera bound both IBSA and IALA, the anti-BSA activity was usually, but not always, greater than the anti-ALA activity. 7. No shared antigenicity was detected between IBSA and ovalbumin, insulin, protamine, diptheria, and tetanus toxoid, pertussis vaccine, poliomyelitis vaccine, and influenza vaccine. The apparent inhibiting effects of unlabeled bovine gamma-globulin and ALA on IBSA binding were probably due to trace amounts of BSA in these protein preparations. 8. BSA, ovalbumin, and bovine gamma-globulin had no detectable shared anti-genicity with IALA. 9. Positive skin tests to milk or BSA did not correlate with the anti-BSA levels measured in the serum. 10. The incidence of persons with anti-BSA and anti-ALA was comparable among the "patient" and "well" populations. Ten of the 31 sera with the greatest capacity to bind IBSA were from the "well" population, the remaining 21 sera were from children with a variety of disease states.

scholarly journals HISTOLOGICAL AND SEROLOGICAL SEQUENCES IN EXPERIMENTAL HYPERSENSITIVITY

1947 ◽  
Vol 85 (6) ◽  
pp. 571-590 ◽  
Author(s):  
Clinton Van Zandt Hawn ◽  
Charles A. Janeway
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Gamma Globulin ◽  
Horse Serum ◽  
Protein Solutions ◽  
Pathological Lesions ◽  
Intravenous Injections ◽  
Foreign Proteins ◽  
Antigen Antibody

1. Groups of normal rabbits were given, single intravenous injections of foreign proteins in doses of 1 gm. per kilo, bled at regular intervals for serologic studies, and sacrificed after varying lengths of time for pathological studies. The protein solutions used were of crystallized bovine serum albumin, bovine serum gamma globulin, and bovine serum. The experiments were planned, first, to correlate the sequence of pathological and immunological changes, and second, to compare the responses to two chemically and immunologically distinct plasma protein fractions and to the whole serum of the same species. 2. (a) The principal pathological lesions in rabbits given bovine serum were similar to those which have been previously observed following, the injection of horse serum and were characterized by widely dispersed but segmental acute inflammatory lesions of the arteries. These lesions were at their height 2 weeks after injection and showed marked repair at 4 weeks. (b) Crystallized bovine serum albumin produced lesions almost exclusively confined to the arteries which were at their height at 2 weeks, were healing at 3, and healed by 4 weeks. The lesions were less numerous and less intense than in animals given whole serum and were only found in some of the animals. (c) Bovine serum gamma globulin elicited quite different histologic sequences. The most striking lesions involved the glomeruli of the kidneys, and to a lesser degree, the heart. Lesions in the liver and joints were present but less conspicuous, and arterial lesions were rare and slight in degree. The lesions not only differed from those in rabbits given albumin in distribution but in timing, since they were most widespread and acute at 1 week and were healing at 2 weeks after injection. Moreover, lesions were observed in almost every animal. 3. Results of immunological studies were consistent with the interpretation that the pathological lesions were due to an antigen-antibody reaction in the tissues, as shown by the following: (a) Acute lesions were only observed when antigen was present and before antibody appeared in the circulation. (b) Healing of lesions was only observed (with one exception) when antigen had almost or completely disappeared from the circulation, usually with the appearance of antibody. (c) There was a correlation between the rapidity of evolution of the lesions and the rapidity with which the antigen disappeared from the circulation. (d) There was a rough correlation between the proportion of animals showing lesions and the proportion developing antibodies after the injection of a particular protein solution.

Antigenic Determinants of Bovine Serum Albumin

2001 ◽  
Vol 126 (3) ◽  
pp. 188-195 ◽  
Author(s):  
Barbara Beretta ◽  
Amedeo Conti ◽  
Alessandro Fiocchi ◽  
Antonella Gaiaschi ◽  
Corrado L. Galli ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Antigenic Determinants

Visualization of native chromatin fibers in different embedding media

1978 ◽  
Vol 36 (2) ◽  
pp. 222-223
Author(s):  
Sebastian Muñoz-Guerra ◽  
Juan A. Subirana
Keyword(s):  
Fine Structure ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Water Soluble ◽  
Critical Importance ◽  
Thin Sections ◽  
Glycol Methacrylate ◽  
Embedding Medium ◽  
Holothuria Polii

The influence of the embedding medium is of critical importance in the observation of nucleoprotein structures in thin sections. The use of conventional epoxi media requires dehydratation, impregnation and subsequent polymerization, which may produce drastic alterations in the fine structure of chromatin. The use of water soluble embedding media may lead to improvements in the preservation of ultrastructure. In this paper we explore the differences which may be detected in nucleohistone fibers embedded in different media.Spermatozoa from Holothuria polii were subjected to moderate lysis in either 0.15M NaCl, ImM Tris-HCl, 0.4mM CaCl2, pH 8.0 (a) or 0.25M sucrose, 0.4mM CaCl2 (b) and then fixed by addition of 2 % glutaralde- hyde. Embedding was carried out as described in the literature in the following media: araldite-epon mixture (AREPO), glycol methacrylate (GMA, 1), hydroxipropyl methacrylate (HPMA, 2) and bovine serum albumin (BSA, 3) or histones.

X-ray diffraction of oriented clays in small quantities (0·1 mg)

Clay Minerals ◽  
1982 ◽  
Vol 17 (2) ◽  
pp. 259-262 ◽  
Author(s):  
A. Meunier ◽  
B. Velde
Keyword(s):  
Thin Section ◽  
Mineral Formation ◽  
X Ray Diffraction ◽  
Oriented Sample ◽  
Thin Sections ◽  
X Ray ◽  
Precise Identification ◽  
Ideal Method ◽  
Subsequent Identification ◽  
The Ideal

Precise identification of clay minerals found in granular rocks has always posed a great problem to the clay petrographer. Even if it is possible to locate the position of authigenic clay mineral formation in a thin section, subsequent identification of this same material by X-ray diffractometry is usually very difficult. Attempts have been made using selected-area radiation of thin sections (Pawluck & Dumanski, 1973; Wicks & Zussman, 1975; Wilson & Clark, 1978) but the area analysed remains relatively large, i.e. of the order of several mm2. The other solution is micro-picking of material from a thin section and subsequent identification by Debye-Scherrer camera methods (Wallace, 1955; Rickwood, 1977). This method, however, does not allow preferred orientation, and thus precise identification, of many clay species. The ideal method is to combine micro-picking from thin sections from areas of several hundreds of square microns with an oriented sample preparation, which can then be treated in the traditional way (glycolation, heating, etc.) for characterization by X-ray diffractometry.

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